Characterization of the Specific Binding Between Aptamers and Cytochrome <i>c</i> With Pressure‐Assisted Capillary Electrophoresis Frontal Analysis
Bibliographic record
Abstract
ABSTRACT Cytochrome c (cyt c ) is a heme protein located in the mitochondrial intermembrane space. Because the release of cyt c is a highly specific event in apoptotic signaling, it can serve as an apoptosis‐related marker. To date, three frequently used aptamers for cyt c (Apt40, Apt61, and Apt76) have been selected and applied in the field of sensing. The response of these aptamers is not clear, partly because of their weak affinity and nonspecific binding inherent to the system. In this study, pressure‐assisted capillary electrophoresis frontal analysis (PACE‐FA) was used to characterize the interactions between the aptamers and cyt c , and an electrophoretic mobility‐based correction was introduced to obtain accurate binding constants. A nonlinear curve‐fitting approach was used for evaluating specific binding interactions in the presence of nonspecific binding. Apt76 was found to bind specifically to cyt c , exhibiting the highest binding constant (1.53 × 10 6 M −1 ), and all three aptamers interacted with cyt c at 1:1 stoichiometry. Fluorescence titrations were performed to verify the effectiveness of the reference‐free PACE‐FA method. This study demonstrates that specific binding between biomolecules has different characteristics compared to nonspecific binding and that the PACE‐FA method can be widely used in the evaluation of biological macromolecular interactions.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".